Pleuromutilin derivatives having antibacterial activity

ABSTRACT

A compound selected from 14—O—[(cycloalkyl-sulfanyl)acetyl]mutilins; 14—O—[(cycloalkyl-alkyl-sulfanyl)acetyl] mutilins; 14—O—[(cycloalkoxy)acetyl] mutilins; and 14—O—[(cycloalkyl-alkoxy)acetyl] mutilins and its use as a pharmaceutical.

[0001] The present invention relates to antibacterials; morespecifically to mutilins. Pleuromutilins are naturally occurringantibiotics which have antimycoplasmal activity and modest antibacterialactivity. We have found mutilins having the principal ring structure ofnaturally occurring pleuromutilins which have improved antimicrobial,e.g. antibacterial activity.

[0002] In one aspect the present invention provides a compound selectedfrom 14—O—[(cycloalkyl-sulfanyl)acetyl]mutilins;14—O—[(cycloalkyl-alkyl-sulfanyl)acetyl] mutilins;14—O—[(cycloalkoxy)acetyl] mutilins; and14—O—[cycloalkyl-alkoxy)acetyl]mutilins; such as 14—O—[(aminocycloalkyl-sulfanyl)acetyl]mutilins; 14—O—[(aminocycloalkyl-alkyl-sulfanyl)acetyl] mutilins;14—O—[(aminocycloalkoxy)acetyl] mutilins; and14—O—[(aminocycloalkyl-alkoxy)acetyl] mutilins; preferably14—O—[(aminocycloalkyl-sulfanyl)acetyl]mutilins; e.g. cycloalkyl ispreferably (C₃₋₁₂)cycloalkyl; cycloalkoxy is preferably(C₃₋₁₂)cycloalkoxy; alkyl is preferably (C₁)alkyl; and alkoxy ispreferably (C₁₋₄)alkoxy.

[0003] In another aspect the present invention provides a compound offormula

[0004] wherein

[0005] R is hydrogen;

[0006] R₁ is hydrogen or a group of formula

[0007] wherein

[0008] X is sulphur, oxygen or NR,₁₀, wherein R₁₀, is hydrogen or alkyl;and

[0009] R₉ is amino, alkyl, aryl or heterocyclyl; and, if X is oxygen, R₉is additionally hydrogen;

[0010] Y is sulphur or oxygen;

[0011] R₂ is hydrogen or one or more substituents, e.g. includingsubstituents such as conventional in organic, e.g. (pleuro)mutilin,chemistry;

[0012] R₄ is hydrogen or alkyl;

[0013] R₅ is hydrogen or alkyl;

[0014] R₃ and R₃′ are hydrogen, deuterium, or halogen;

[0015] R₆, R₇ and R₈ are hydrogen or deuterium;

[0016] m is a number selected from 0 to 4;

[0017] n is a number selected from 0 to 10; and

[0018] p is a number selected from 0 to 10; with the proviso that n plusp are at least 1 and

[0019] preferably less than 13.

[0020] In formula I, preferably

[0021] R is hydrogen;

[0022] R₁ is hydrogen or a group of the formula −C(=X)R₉,

[0023] wherein X is oxygen; and

[0024] R₉, is alkyl, e.g. (C₁₋₈)alkyl, such as (C₁₋₄)alkyl,unsubstituted or substituted alkyl, e.g. substituted by groups which areconventional in organic, e.g. pleuromutilin, chemistry, such as one ormore amino; e.g., if R₉ is alkyl substituted by amino, R₉ is preferablythe residue of an amino acid, e.g. valine, e.g. said residue includesthat part of an amino acid which remains if the carboxylic group issplit off;

[0025] Y is sulphur;

[0026] R₂ is hydrogen;

[0027] R₄, R₅, R₃, R′₃, R₆, R₇, and R₈ are hydrogen;

[0028] m is o;

[0029] n is 3 or4;

[0030] p is 0 or 1; and

[0031] p plus n is 3 or4.

[0032] In another aspect, the invention provides a compound of theformula l_(p)

[0033] wherein R_(1p) is hydrogen or the residue of an amino acid; e.g.valyl; e.g. R_(1p) is a group of formula —CO—R_(9p) wherein R_(9p) isthe residue of an amino acid which remains if the carboxylic group issplit off.

[0034] In formula l_(p) a group —NH—R_(1p) may be in any position of thecyclohexyl ring system and is preferably in position 2, 3, or 4. Aminoacid in the meaning of Rap includes any amino acid, preferably valine;and R_(9p) is preferably a group —CH(NH₂)—CH(CH₃)₂. The amine group insaid amino acid residue may be unprotected or protected, e.g. byappropriate amino acid protection groups, e.g. such as conventional, forexample tert.butoxycarbonyl; and is preferably unprotected.

[0035] In another aspect, the present invention provides a compoundwhich is:

[0036]14—O—[(aminocyclohexan-2-yl-sulfanyl)acetyl]mutilin,

[0037]14—O—[(aminocyclohexan-3-yl-sulfanyl)acetyl]mutilin,

[0038]14—O—[(aminocyclohexan-4-yl-sulfanyl)acetyl]mutilin,

[0039]14—O—[(N-valyl-aminocyclohexan-2-yl)sulfanyl)-acetyl]-mutilin,

[0040]14—O—[(N-valyl-aminocyclohexan-3-yl)sulfanyl)-acetyl]-mutilin, or

[0041]14—O—[(N-valyl-aminocyclohexan-4-yl)sulfanyl)-acetyl]-mutilin;including

[0042]14—O—[(N-(R)-valyl-(R)-aminocyclohexan-2(R)-yl)sulfanyl)-acetyl]-mutilin,

[0043]14—O—[(N-(R)-valyl-(R)-aminocyclohexan-2(S)-yl)sulfanyl)-acetyl]-mutilin,

[0044]14—O—[(N-(R)-valyl-(R)-aminocyclohexan-3(R)-yl)sulfanyl)-acetyl]-mutilin,

[0045]14—O—[(N-(R)-valyl-(R)-aminocyclohexan-3(S)-yl)sulfanyl)-acetyl]-mutilin;

[0046]14—O—[(N-(R)-valyl-cis-aminocyclohexan-4-yl)sulfanyl)-acetyl]-mutilin,and

[0047]14—O—[(N-(R)-valyl-trans-aminocyclohexan-4-yl)sulfanyl)-acetyl]-mutilin,e.g., in free (base) form or in the form of a salt, such as ahydrochloride.

[0048] Preferred compounds include:14—O—[(aminocyclohexan-4-yl-sulfanyl)acetyl]mutilin and14—O—[(N-valyl-aminocyclohexan-4-yl)sulfanyl)-acetyl]-mutilin. Morepreferred is14—O—[(N-valyl-aminocyclohexan-4-yl)sulfanyl)-acetyl]-mutilin.

[0049] A compound provided by the present invention is hereinafterdesignated as “A compound of the present invention”. The presentinvention includes a compound of the present invention, e.g. including acompound of formulae I and lp, in free (base) form and in the form of asalt, e.g. in the form of a solvate.

[0050] In another aspect, the present invention provides a compound ofthe present invention in the form of a salt, e.g. and in the form of asalt and in the form of a solvate, or in the form of a solvate.

[0051] A salt of a compound of the present invention includes apharmaceutically acceptable salt, e.g. including a metal salt or an acidaddition salt. Metal salts include for example alkali or earth alkalisalts; acid addition salts include salts of a compound of the presentinvention with an acid, e.g. hydrogen fumaric acid, fumaric acid,naphthalin-1,5-sulfonic acid, hydrochloric acid, and deuterochloricacid; preferably hydrochloric acid or deuterochloric acid.

[0052] A compound of the present invention in free form may be convertedinto a corresponding compound in the form of a salt; and vice versa. Acompound of the present invention in free form or in the form of a saltand in the form of a solvate may be converted into a correspondingcompound in free form or in the form of a salt in unsolvated form; andvice versa.

[0053] A compound of the present invention may exist in the form ofisomers and mixtures thereof; e.g. a compound of the present inventionmay contain asymmetric carbon atoms and may thus exist in the form ofdiastereoisomeres and mixtures thereof. For example, in a compound offormula lp, wherein the group —NH—R_(1p) is in position 2 or 3 of thecyclohexyl ring, the carbon atom of the cyclohexyl ring which isattached to the side chain of the mutilin ring and the carbon atom ofthe cyclohexyl ring to which the group —NH—R_(1p) is attached are bothasymmetric carbon atoms. A compound of formula l_(p) wherein the group—NH—R_(1p) is in position 2 or 3 of the cyclohexyl ring may thus existin (R) and (S) configurations in respect to both of these carbon atoms.For example, if R_(1p) is the residue of an amino acid, that amino acidmay comprise asymmetric carbon atoms. E.g., if R_(1p) is valyl, thecarbon atom to which the amine group of said valyl is attached is anasymmetric carbon atom. A compound of formula l_(p) wherein R_(1p) isvalyl may thus exist in (R) and in (S) configurations in respect to saidvalyl carbon atom.

[0054] Isomeric or diastereoisomeric mixtures may be separated asappropriate, e.g. according to conventional methods, to obtain pureisomers or diastereoismers, respectively. The present invention includesa compound of the present invention in any isomeric and diasteroisomericform and in any isomeric and diastereoisomeric mixture. Preferably theconfiguration in the mutilin ring of a compound of formula I is the sameas in a naturally produced pleuromutilin.

[0055] A compound of the present invention may be obtained asappropriate, e.g. according to, e.g. analogously, to any conventionalmethod. E.g., 14-O-[(cycloalkyl-sulfanyl)acetyl]mutilins;14—O—[(cycloalkyl-alkyl-sulfanyl)acetyl] mutilins;14—O—[(cycloalkoxy)acetyl] mutilins; and14—O—[(cycloalkyl-alkoxy)acetyl] mutilins of the present invention maybe prepared by reacting a 14—O—[(mercapto)acetyl]mutilin or a14—O—[(hydroxy)acetyl] mutilin, respectively, with a hydroxycyclalkyl,or a hydroxyalkyl-cycloalkyl, respectively, in an activated form, e.g.in the form of an ester with a sulfonic acid, and isolating a compoundof the present invention from the reaction mixture obtained.

[0056] Any compound of the present invention and any intermediate in thepreparation of a compound of the present invention may be obtained asappropriate, e.g. according, such as analogously, to a conventionalmethod, e.g. or as specified herein, including the examples. A compoundof formula I or I_(p) may be obtained. e.a. according. e.q. analoqously.to a process for the preparation of14—O—[(cycloalkyl-sulfanyl)acetyl]mutilins;14—O—[(cycloalkyl-alkyl-sulfanyl)acetyl] mutilins; 14—O—[(cycloalkoxy)acetyl] mutilins; and14—O—[(cycloalkyl-alkoxy)acetyl]mutilins.

[0057] In another aspect, the present invention provides a process forthe production of a compound of formula I comprising the steps

[0058] a. reacting a compound of formula II

[0059] wherein Y, R₃, R′₃, R₄, and R₅ are as defined above and R₆, R₇,and R₈ are hydrogen, with a compound of formula III

[0060] wherein R, R₁, R₂, m, n, and p are as defined above, in anactivated form, e.g. in the form of an ester with 4-toulenesulfonic acid(tosylate), or of an ester with methylsulfonic acid (mesylate), toobtain a compound of formula I; wherein R₆, R₇ and R₈ are hydrogen andY, R₁, R, R₂, R₃, R′₃, R₄, R₆, m, n, and p are as defined above, and, ifdesired, b. introducing deuterium into a compound of formula I obtainedin step a, to obtain a compound of formula I, wherein Y, R₁, R, R₂, R₃,R′₃, R₄, R₅, m, n, and p are as defined above and R₆, R₇, and R₈ aredeuterium.

[0061] A compound of formula II is known or may be obtained according,e.g. analogously, to a conventional method. A compound of formula IIImay be obtained as appropriate, e.g. according, e.g. analogously, to aconventional process. A compound of formula III may preferably beprepared according to the following process, e.g. or as described in theexamples:

[0062] A dihydroxycycloalkyl or a (hydroxyalkyl)(hydroxy)cycloalkylrespectively, wherein R₂, m, p, and n are as defined above; may bereacted in solvent which is inert under the reaction conditions with4-toluenesulfonic or methanesulfonic acid anhydride to obtain acorresponding di-tosyl/mesyl-oxycycloalkyl, or(tosyl/mesyl)oxyalkyl)(tosyl/mesyloxy) cycloalkyl respectively; whichdi-tosyl/mesyl-compound is further reacted with sodium azide to obtain acorresponding (tosyl/mesyloxy)(azido)cycloalkyl, or a(tosyl/mesyloxyalkyl)(azido)cycloalkyl, respectively. In a(tosyl/mesyloxy)(azido)-compound obtained the azido group is reduced,e.g. catalytically hydrogenated, to obtain the corresponding(tosyl/mesyloxy)(amino)cycloalkyl, or(tosyl/mesyloxyalkyl)(amino)cycloalkyl, respectively; which is acompound of formula 111, wherein the hydroxy group istosylated/mesylated; wherein R and R₁ are hydrogen and wherein R₂, m, pand, n are as defined above. If desired, the amine group obtained byreduction of the azido group may be reacted with R₉—C(=X)OH, wherein R₉and X are as defined above, in an activated form, e.g. if X is oxygenR1—C(=X)OH may be in the form of an anhydride, halogenide; to obtain acompound of formula III, wherein R is hydrogen; R₁ is a group of formula—C(=X)R9, wherein X and R₉ are as defined above; and wherein R₂, m, p,and n are as defined above.

[0063] Replacement of hydrogen atoms in a compound of formula 1, e.g. inthe form of a salt; by deuterium atoms may be carried out asappropriate, e.g. according, e.g. analogously, to a conventional method,e.g. or according to a method described herein; e.g. by treatment of acompound of formula 1, e.g. including a compound of formula Ip; withdeuterochloric acid (DCI) in appropriate solvent (system) and isolationof a compound of formula I, e.g. in the form of a salt, wherein hydrogenatoms, e.g. in the meaning of R₆, R₇, and R₈ are replaced by deuteriumatoms.

[0064] The production of a compound of formula I, wherein R₃ and R′₃ aredeuterium or halogen may be carried out as appropriate, e.g. according,e.g. analogously, to a conventional method, e.g. via treatment of acompound of formula V

[0065] wherein the carbon atoms carrying R₃ and R′₃, which both arehydrogen, together form a double bond and which is a known compound,with deuterium or with halogen, e.g. with F₂, Cl₂, or, Br₂, to obtain acompound of formula V, wherein R₃ and R′₃ are deuterium or halogen; andfurther reacting a compound of formula V, wherein R₃ and R′₃ aredeuterium or halogen as appropriate, e.g. according, e.g. analogously,to a conventional method, to obtain a compound of formula II, wherein,R₃ and R′₃ are deuterium or halogen and R₆, R₇, and R₈ are hydrogen.

[0066] Preferably a compound of formula II may be obtained from acompound of formula V by reacting a compound of formula V with acompound of formula III

[0067] wherein Y, R₄, and R₅ are as defined above and Hal is halogen,preferably bromo or chloro. A compound of formula III is known or may beobtained as appropriate, e.g. according, e.g. analogously, to aconventional method.

[0068] The compounds of the present invention, e.g. including a compoundof formulae I and I_(p), hereinafter designated as “active compound(s)of the present invention” exhibit pharmacological activity and aretherefore useful as pharmaceuticals. For example, the active compoundsof the present invention show antimicrobial, e.g. antibacterial,activity against gram positive bacteria, such as Staphylococci, e.g.Staphylococcus aureus, Streptococci, e.g. Streptococcus pyogenes,Streptococcus pneumoniae, Enterococci, e.g. Enterococcus faecium, aswell as against mycoplasms, Chlamydia, and obligatory anaerobes, e.g.Bacteroides fragilis; in vitro in the Agar Dilution Test orMicrodilution Test according to National Commitee for ClinicalLaboratory Standards (NCCLS) 1997, Document M7-A4 Vol.17, No. 2:“Methods for dilution Antimicrobial Susceptibility Tests for Bacteriathat Grow Aerobically—Fourth Edition, Approved Standard”; and in theAnaerobic Bacteria TEST according to National Committee for ClinicalLaboratory Standards (NCCLS) VOL. 13, No. 26; M11-A4, Methods forAntimicrobal Susceptibility Testing of Anaerobic Bacteria; ApprovedStandard; Fourth Edition (1997). For example, the compound of example 1shows in vitro in the above indicated Agar Dilution Test and/orMicrodilution Test against bacterial strains as mentioned above MICvalues of 0.01 to 1.0 μg/mL.

[0069] In another aspect, the present invention provides a compound ofthe present invention for use as a pharmaceutical, preferably as anantimicrobial, such as an antibiotic, e.g. and an anti-anaerobic.

[0070] In a further aspect, the present invention provides a compound ofthe present invention for use in the preparation of a medicament for thetreatment of microbial diseases, for example of diseases caused bybacteria, e.g. selected from Staphylococci, Streptococci, Enterococci;e.g. and of diseases caused by mycoplasms, Chlamydia, and obligatoryanaerobes.

[0071] In a further aspect, the present invention provides a method oftreatment of microbial diseases which comprises administering to asubject in need of such treatment an effective amount of a compound ofthe present invention e.g. in the form of a pharmaceutical composition.

[0072] For antimicrobial treatment, the appropriate dosage will, ofcourse, vary depending upon, for example, the active compound of thepresent invention employed, the host, the mode of administration, andthe nature and severity of the conditions being treated. However, ingeneral, for satisfactory results in larger mammals, for example humans,an indicated daily dosage is in the range from about 0.5 to 3 g, of anactive compound of the present invention conveniently administered, forexample, in divided doses up to four times a day.

[0073] An active compound of the present invention may be administeredby any conventional route, preferably orally, e.g. in form of tablets,powders, capsules, or suspensions; e.g. including non-resorbable oralformulations; or parenterally, e.g. in the form of injectable solutionsor suspensions; or topically, e.g. in the form of nasal sprays, bodysolutions, creams, or eye drops. The active compounds of the presentinvention may be administered in analogous manner, e.g. in similar dosesand for similar indications, as ervthromycin(s), tetracycline(s).Surprisingly the active compounds of the present invention show alsoactivity against strains which are resistant against erythromycin(s),tetracycline(s).

[0074] The active compounds of the present invention may be administeredin the form of a pharmaceutically acceptable salt, e.g. an acid additionsalt or metal salt; or in free form; optionally in the form of asolvate. The active compounds of the present invention in the form of asalt exhibit the same order of activity as the active compounds of thepresent invention in free form. The active compounds of the presentinvention may be administered in the form of pharmaceuticalcompositions.

[0075] In another aspect he present invention provides a pharmaceuticalcomposition comprising a compound of the present invention, e.g. in freeform or in the form of a pharmaceutically acceptable salt; e.g. and/orin the form of a solvate; in association with at least onepharmaceutical carrier or diluent.

[0076] Such compositions may be manufactured according, e.g.analogously, to a conventional method. Unit dosage forms may contain,for e.g., about 100 mg to about 1 g.

[0077] The active compounds of the present invention are additionallysuitable as veterinary agents, e.g. veterinary active compounds, e.g. inthe prophylaxis and in the treatment of microbial, e.g. bacterialdiseases, in animals, such as fowl, pigs, and calves; e.g. and fordiluting fluids for artificial insemination and for egg-dippingtechniques.

[0078] In another aspect, the present invention provides a compound ofthe present invention for use as a veterinary agent.

[0079] In a further aspect, the present invention provides a compound ofthe present invention for the preparation of a veterinary compositionwhich is useful as a veterinary agent.

[0080] The present invention further provides a veterinary method forthe prophylaxis and in the treatment of microbial, e.g. bacterialdiseases, which comprises administering to a subject in need of suchtreatment an effective amount of a compound of the present invention,e.g. in the form of a veterinary composition.

[0081] For use of the active compounds of the present invention as aveterinary agent, the dosage will of course vary depending upon the sizeand age of the animal and the effect desired. For example, forprophylactic treatment relatively low doses would be administered over alonger time period, e.g. 1 to 3 weeks. Preferred doses in drinking waterare from 0.0125 to 0.05 weight by volume, particularly 0.0125 to 0.025;and in foodstuffs from 20 to 400 g/metric ton, preferably 20 to 200g/metric ton. It is preferred to administer the active compounds of thepresent invention as a veterinary agent to hens in drinking water, topigs in foodstuff, and to calves orally or parenterally, e.g. in theform of oral or parenteral preparations.

[0082] In the following examples, which illustrate the invention,references to temperature are in degrees Celsius.

[0083] The following abbreviations are used:

[0084] DCCI dicyclohexylcarbodiimide

[0085] BOC tert.butoxycarbonyl

[0086] DMF dimethylformamide.

[0087] The numbering of the mutilin ring system referred to in theexamples is given in the following formula:

[0088] 14—O—[(3-(R)-amino-2-methyl-propanecarbonylamino)-cyclohexane-1-yl)-sulfanyl)-acetyl]-mutilin(=14—O—[(N-(R)-valyl-(R)-aminocyclohexan-3(R)-yl)sulfanyl)-acetyl]-mutilin)A. 1,3-bis-(4-Toluene-sulfonyloxy)-cyclohexane A solution of 11.6 a of1.3-dihydroxvcyclohexane. 6.52 g of 4-toluenesulfonic acid anhydride and40.4 g of N-methylmorpholine in 200 mL of methylenechloride is stirredfor ca. 24 hours at room temperature. The reaction mixture obtained ispoured onto 1 N HCI and the mixture obtained is extracted withmethylenechloride. The organic phase obtained is dried and the solventis evaporated off. 38.5 g of 1,3-bis-(4-toluene-sulfonyloxy)-cyclohexaneare obtained. B. 1-Toluenesulfonyloxy-3-azido-cyclohexane

[0089] To a solution of 3.9 of 1,3g-bis-(4-toluene-sulfonyloxy)-cyclohexane in 100 mL of DMF, 0.55 g ofsodium azide is added in portions. The reaction mixture obtained isheated to ca. 80° for ca. 2 hours, the solvent is removed under vacuum,and the residue obtained is dissolved in 200 mL of methylene chlorideand subjected to chromatography. 1.2 g of1-toluenesulfonyloxy-3-azido-cyclohexane are obtained. H¹-NMR(CDCl₃):Mixture of diastereoisomeres: 7.8,7.3(2xd, 4H, arom.H), 4.8,4.4(2xm,1H,CHO), 3.2,3.7(2xm, 1H,CHN), 2.4(s,3H,arom.CH₃),1.2-1.8(m,8H,cyclohexyl).

[0090] C.1-Toluenesulfonyloxy-3-(3(R)-tert.butyloxycarbonylamino-2-methyl-propanecarbonylamino)-cyclohexane

[0091] A solution of 2.75 g of 1-toluenesulfonyloxy-3-azido-cyclohexanein 50 mL of ethyl acetate is hydrogenated in the presence of a catalyticamount of palladium on carbon (10%). The catalyst is filtrated off and2.2 g of N-BOC-(D)-valine and 2 g of DCC are added to the filtrateobtained. The reaction mixture obtained is stirred for ca. 15 hours atroom temperature, filtered and the solvent is evaporated off. Theresidue obtained is subjected to chromatography. 1.4 g of 1-toluenesulfonyloxy-3-(3(R)-tert.butyloxycarbonylamino-2-methyl-propanecarbonyl-amino)-cyclohexane areobtained. H¹-NMR(CDCl₃): Mixture of diastereoisomeres:7.8,7.3(2xm, 4H,arom.H), 6.5(m,1H,NH), 4.8,4,43(2xm, 1H,CHO), 3.85,3.55(2xm,1H, CHN),3.62,3.7(2xm,1H,NCHCO), 2.4(s,3H,arom.CH₃),1.82(m,1H,CHC(CH₃)₂),1.38(b,9H,BOC), 0.78(m,6H, CHC(CH₃)₂).

[0092] D.14-O-[(3-(3-(R)-Amino-2-methyl-propanecarbonylamino)-cyclohexane-1-yl)-sulfanyl-acetyl]-mutilin

[0093] To a solution of 1.75 g of 14-mercaptoacetyl-mutilin and 70 mg ofsodium in 100 mL of ethanol, 1.4 g of1-toluenesulfonyloxy-3-(3(R)-tert.butyloxycarbonylamino-2-methyl-propanecarbonylamino)-cyclohexaneand 0.7 mL of N-methylmorpholine are added. The mixture obtained isheated for ca 8 hours at ca. 90° C. The mixture obtained is poured ontobrine, the mixture obtained is extracted with ethyl acetate, and theorganic phase obtained is evaporated off. The residue obtained istreated with a mixture of trifluoroacetic acid/methylenechloride 1:1 andsubjected to chromatography.14-O-[(3-(3-(R)-amino-2-methyl-propanecarbonylamino)-cyclohexane-1-yl)-sulfanyl-acetyl]-mutilinis obtained. H¹-NMR(d₆DMSO,350K): Mixture of diastereoisomeres:7.4(d,1H,NH), 6.15(m,1 H,H19), 5.55(d,1H,H14), 5.05(m,2H,H20),4.5(d,1H,H11),3.78(2xd,1H, NCHCO), 3.9(m,1H,NCH), 3.42(t,1H,H11),3.25(m,2H,SCH₂CO),3.2(m,1HCHS), 0.9,0.88 (2xd,6H,(CH₃)₂CH),1.08,1.36(2xs,6H,(CH₃)₁₈, (CH₃)₁₅), 0.65,0.83(2xd,6H, (CH₃),₆, (CH₃)₁₇).¹H-NMR (CDCl₃): ABX-system (ν_(A)=3.15, ν_(B)=3.22, ν_(x)=1.92, 2H, H₂₂,J=15.8Hz, J=8.2Hz)

Example 2

[0094] A. N-(R)-(N-BOC-(R)-Valyl)-2(R)-hydroxy-cyclohexylamine

[0095] A mixture of 1.5 g trans-2-aminocyclohexanol, 2.17 g ofBOC-(R)-valine, 2.06 g of DCC, and 1.01 g of N-methylmorpholine in 40 mLof CH₂Cl₂ is kept for ca. 24 hours at 250. A precipitate (urea) isformed and is filtrated off. The filtrate obtained is extracted withwater, dried, the solvent is evaporated off, and the evaporation residueobtained is subjected to chromatograpy over silica gel. 710 mg ofN-(R)-(N-BOC-(R)-valyl)-2(R)-hydroxy-cyclohexylamine are obtained. H¹NMR (d₆DMSO, mixture of diastereoisomeres): 6.1,6.25(2xd,1H,CONH),5.1,5.2(2xb,1 H,BOC-HN), 3.85(m,1 H,a -H-val), 3.3,3.65(2xm,2H,NCH,OCH).

[0096] B. N-(R)-(N-BOC-(R)-valyl)-2(R)-methansulfonyloxy-cyclohexylamine

[0097] A mixture of 710 mg ofN-(R)-(N-BOC-(R)-valyl)-2(R)-hydroxy-cyclohexylamine, 393 mg ofmethanesulfonic acid anhydride, and 236 mg of N-methyl morpholine in 15mL of CH₂Cl₂ is stirred at 250 for ca. 12 hours. The mixture obtained isextracted with 1N HCl, dried and the solvent is evaporated off. 685 mgof N-(R)-(N-BOC-(R)-valyl)-2(R)-methansulfonyloxy-cyclohexylamine areobtained.

[0098]C.14-O-[(N-(R)-Valyl-(R)-aminocyclohexan-2(S)-yl)sulfanyl)-acetyl]-mutilinin the form of a hydrochloride +(RSR)-Diastereomeres 1:1

[0099] A mixture of 588 mg ofN-(R)-(N-BOC-(R)-valyl)-2(R)-methansulfonyloxy-cyclohexylamine, 591 mgof 22-desoxy-22-pleuromutilin-thiol, and 35 mg of sodium in 10 mL of dryehthanol is stirred for ca. 24 hours at 25° and heated for ca. 2 hoursat 90°. From the mixture obtained the solvent is evaporated off. To theevaporation residue ethyl acetate and water are added, the phasesobtained are separated, and the organic phase is subjected tochromatography over silica gel. 14—O—[(N-(R)-(N-BOC-(R)-valyl)-(R)-aminocyclohexan-2(S)-yl)sulfanyl)-acetyl]-mutilinis obtained and is treated with etheric HCl. 940 mg of 14—O—[(N—R)—valyl—(R)-aminocyclohexan-2(S)-yl)sulfanyl)-acetyl]-mutilin-in theform of a hydrochloride are obtained. H¹NMR(d₆DMSO, 350K, mixture ofdiastereoisomeres): 6.15 (m,1H,H₁₉), 5.56, 5.58 (2xd, 1H,H₁₄), 5.1(m,2H,H₂₀), 4.13(d,1H,OH, J=5.5 Hz), 3.43(t,1H,H1 ,J=5.5 Hz), AB-system(ν_(A) =3.28,ν_(B)=3.23, 2H,H22,J=14.7 Hz), 2.95(m,1H,CHS),2.75(m,1H,CHNH), 0.65, 0.85 (2xd,6H, (CH₃)CH, 6.9 Hz),1.08,1.37(2xs,6H,(CH₃)₁₈, (CH₃)₁₅.

[0100] D. 14-O-[(N-(R)-Valyl-(R)-aminocyclohexan-2(R)-yl)sulfanyl)-acetyl]-mutilin inthe form of a hydrochloride+(RSR)-Diastereomeres 1:1

[0101] The title compound is obtained analogously as described inExample 2, step C), but using the appropriate starting materials, namelyN-(R)-(N-BOC-(R)-valyl)-2(S)-methansulfonyloxy-cyclohexylamine insteadof N-(R)-(N-BOC-(R)-valyl)-2(R)-methansulfonyloxy-cyclohexylamine.H¹NMR(d₆DMSO, 350K, mixture of diastereoisomeres): 8.0(b,3H,NH₃),8.25(m, 1 H,NH), 6.15 (m,1H,H₁₉), 5.56, 5.58 (2xd, 1H,H₁₄), 5.1(m,2H,H₂₀), 3.2-3.5(m, 5H,Hll, NHCH, SCH,CH₂S), 3.6 (m,1H,α-H-val),2.75(m,1H,CHNH), 0.65, 0.85 (2xd,6H, (CH₃)CH, 6.9 Hz), 1.08,1.37(2xs,6H,(CH₃)₁₈, (CH₃)₁₅.

[0102] Example 3

[0103]14-0-[(N-valyl-aminocyclohexan-4-yl)sulfanyl)-acetyl]-mutilin inthe form of a hydrochloride

[0104] A. 14-O-[(Carbamimidoylsulfanyl)acetyl]mutilin-tosylate

[0105] A solution of 15.2 g of thiourea and 106.4 g ofpleuromutilin-22—O—tosylate in 250 mL of acetone is heated under refluxfor 1.5 hours, solvent is removed under reduced pressure, and 100 mL ofhexane is added. A precipitate forms, is filtrated off and dried.14-O-[(carbamimidoylsulfanyl)acetyl]mutilin-tosylate is obtained. ¹H-NMR(CDCl₃): AB-system (ν_(A)=3.7, ν_(B) =3.82, 2H, H₂₂, J=15.8Hz), 7.2 (d,2H, arom.H, J=8 Hz), 7.75 (d, 2H, arom.H, J=8 Hz), 8.4, 9.8 (2xb, 4H,2xNH₂).

[0106] B. 14-Mercapto-acetyl-mutilin

[0107] A solution of 4.7 g of sodium pyrosulfite (Na₂S₂O₅) in 25 mL ofH₂O is added to a solution of 12.2 g of14-O-[(carbamimidoylsulfanyl)acetyl]mutilin-tosylate in a mixture of 20mL of ethanol and 35 mL of H₂O (warmed to ca. 900). 100 mL of CCl₄ areadded to the reaction mixture obtained and the mixture is heated underreflux for ca. 2 hours. The two-phase system obtained is separated, theorganic phase is dried, and the solvent is evaporated off.14-Mercapto-acetyl-mutilin is obtained. ¹H-NMR (CDCI₃): ABX-system(ν_(A)=3.15, ν_(B)=3.22, ν_(x),=1.92, 2H, H₂₂, J=15.8 Hz, J=8.2 Hz).

[0108] C. 14-O-[(4-Amino-cyclohexyl-sulfanyl)acetyl]mutilin 394 mg of14-mercapto-acetyl-mutilin are added under stirring at room temperatureto a solution of 23 mg of sodium in absolute ethanol. The mixtureobtained is treated with 190 mg of4-(N-BOC-amino)cyclohexan-1-yl-p-toluenesulfonate), prepared by reactionof 4-(N-BOC-amino)cyclohexanol with p-toluenesulfonic acid anhydride,and the mixture obtained is refluxed for ca. 2 hours.14-O-[(N-BOC-aminocyclohexan-4-yl)sulfanyl)-acetyl]-mutilin is formedand is isolated from the reaction mixture and is treated withtrifluoroacetic acid in order to split off BOC.14-O-[(4-amino-cyclohexyl-sulfanyl)acetyl]mutilin is obtained. ¹H-NMR(d6-DMSO): 7.9 (b,3H,NH₃), AB-system (ν_(A)=3.23, ν_(B)=3.29, 2H, H₂₂,J=15.2Hz),3.03(m,1H,SCH), 3.10(m,1H,CHN).

[0109] D. 14-0-[(N-valyl-aminocyclohexan-4-yl)sulfanyl)-acetyl]-mutilinin the form of a hydrochloride

[0110] 60 mg of DCCI are added to a mixture of 65 mg of N-BOC-valin and98 mg of 14-O-[(4-amino-cyclohexyl-sulfanyl)acetyl]mutilin in 10 mL ofdichloromethane at room temperature and the mixture obtained is stirredfor several hours at room temperature. A precipitate obtained isfiltrated off and the filtrate obtained is concentrated under reducedpressure. The concentrate obtained is subjected to chromatography.14-O-{{[N-(N-BOC-valyl)-aminocyclohexan-4-yl]sulfanyl}acetyl}-mutilin isobtained and is treated with etheric hydrochloric acid at roomtemperature for ca. 1 hour. From the mixture obtained the solvent isremoved under reduced pressure and the residue obtained is precipitated.14-O-[(N-valyl-aminocyclohexan-4-yl)sulfanyl)-acetyl]-mutilin in theform of a hydrochloride is obtained. ¹H-NMR (d6-DMSO): 8.05 (b,3H,NH₃),8.3(d,1H,NH,J=6.7 Hz), AB-system (ν_(A)=3.23, ν_(B)=3.27,2H, H₂₂,J=15.2Hz),2.98(m,1H,SCH), 3.75(m,1H,a-H, amino acid), 0.9(d,6H,2xCH3,J=5.9 Hz).

1. A compound which is a 14—O—[(cycloalkyl-sulfanyl)acetyl]mutilin;14—O—[(cycloalkyl-alkyl-sulfanyl)acetyl] mutilin;14—O—[(cycloalkoxy)acetyl] mutilin; or 14—O—[(cycloalkyl-alkoxy)acetyl]mutilin.
 2. A compound of formula I

wherein R is hydrogen; R₁ is hydrogen or a group of formula

wherein X is sulphur, oxygen, or NR₁₀, wherein R₁₀ is hydrogen or alkyl;and R₉ is amino, alkyl, aryl, or heterocyclyl; and, if X is oxygen, R₉is additionally hydrogen; Y is sulphur or oxygen; R₂ is hydrogen or oneor more substituents; R₄ is hydrogen or alky; R₅ is hydrogen or alkyglR₃ and R₃′ are hydrogen, deuterium, or halogen; R₆, R₇, and R₈ arehydrogen or deuterium; m is a number selected from 0 to 4; n is a numberselected from 0 to 10; and p is a number selected from 0 to 10; with theproviso that n plus p are at least
 1. 3. A compound of the formula I_(p)

wherein R_(1P) is hydrogen or the residue of an amino acid.
 4. Acompound which is 14—O—[(aminocyclohexan-2-yl-sulfanyl)acetyl]mutilin,14—O—[(aminocyclohexan-3-yl-sulfanyl)acetyl]mutilin,14—O—[(N-valyl-aminocyclohexan-2-yl)sulfanyl)-acetyl]-mutilin, or14—O—[(N-valyl-aminocyclohexan-3-yl)sulfanyl)-acetyl]-mutilin.
 5. Acompound which is 14—O—[(aminocyclohexan-4-yl-sulfanyl)acetyl]mutilin,or 14—O—[(N-valyl-aminocyclohexan-4-yl)sulfanyl)-acetyl]-mutilin.
 6. Acompound of claim 1 in the form of a salt.
 7. A pharmaceuticalcomposition comprising an effective amount of a compound of claim 1 andat least one pharmaceutical carrier or diluent therefor.
 8. A method oftreatment of microbial diseases which comprises administering to asubject in need of such treatment an effective amount of a compound ofclaim
 1. 9. A method of claim 8 wherein the subject is a non-humananimal.